CAJ | 학술논문

[目的]以湖北白猪肝脏作为试验材料,克隆出湖北白猪胰岛素生长因子（IGF-1）基因,并对其序列进行分析。[方法]采用Trizol法从湖北白猪的肝脏中提取总RNA,将其作为RT反应的模板,用P2（5＇-CAGGTAACTCGTGCAGAGCAAAGGA-3＇）引物合成IGF-1基因cDNA第一链,以其产物为模板,再以P1（5＇-CCCATCTCCCTGGATTTCTTTTTG-3＇）和P2为上下游引物扩增到大小约为607bp的产物,并将其克隆至pCRII载体上,经蓝白斑筛选、酶切、测序。[结果]经筛选、酶切、序列分析,表明该片段为IGF-1基因的cDNA克隆,它由607个核苷酸组成,其中1～145位是5＇端非翻译区,从第146位ATG起始密码子开始至第538位TAG终止密码子,539～607位是3＇端非翻译区,包含一完整的ORF,该ORF共有393核苷酸,编码一个推断由130个氨基酸组成的多肽。与GenBank中的序列比较,该序列与Muller等报道的猪IGF-1基因编码序列完全同源。[结论]成功克隆了湖北白猪IGF-1基因,并对其进行序列分析,证实了IGF-1基因的高度保守性,为采用转基因技术对湖北白猪进行育种奠定了基础和提供了技术依据。
[목적]이호북백저간장작위시험재료,극륭출호북백저이도소생장인자（IGF-1）기인,병대기서렬진행분석。[방법]채용Trizol법종호북백저적간장중제취총RNA,장기작위RT반응적모판,용P2（5＇-CAGGTAACTCGTGCAGAGCAAAGGA-3＇）인물합성IGF-1기인cDNA제일련,이기산물위모판,재이P1（5＇-CCCATCTCCCTGGATTTCTTTTTG-3＇）화P2위상하유인물확증도대소약위607bp적산물,병장기극륭지pCRII재체상,경람백반사선、매절、측서。[결과]경사선、매절、서렬분석,표명해편단위IGF-1기인적cDNA극륭,타유607개핵감산조성,기중1～145위시5＇단비번역구,종제146위ATG기시밀마자개시지제538위TAG종지밀마자,539～607위시3＇단비번역구,포함일완정적ORF,해ORF공유393핵감산,편마일개추단유130개안기산조성적다태。여GenBank중적서렬비교,해서렬여Muller등보도적저IGF-1기인편마서렬완전동원。[결론]성공극륭료호북백저IGF-1기인,병대기진행서렬분석,증실료IGF-1기인적고도보수성,위채용전기인기술대호북백저진행육충전정료기출화제공료기술의거。
[Objective] The study aimed at cloning and analyzing the insulin-like growth factor-1 （IGF-1） gene from liver of Hubei white swine. [Method] The total RNA was extracted by using Trizol from the liver of Hubei white swine and used as template to amplify IGF-1 gene cDNA by RT-PCR. The cDNA product was cloned into pCRII vector, screened with blue-white colonies, digested with double enzymes and sequenced. [Result] The sequencing result indicated that the IGF-1 gene consisted of 607 nucleotides, containing 5＇-untranslated region at nucleotides 1-145, a complete ORF at nucleotides 146-538 encoding 130 amino acids, and 3＇-untranslated region at nucleotides 539-607. It shared 100% homology with the porcine IGF-1 gene reported by Muller et al. [Conclusion] The successful cloning and sequencing of the Hubei white swine IGF-1 gene confirmed that IGF-I gene was highly conserved, which provided technical basis for the use of transgenic technology for breeding of Hubei white swine.